The E.Z.N.A^® Tissue RNA Kit provides an easy solution for RNA isolation from difficult to lyse samples such as skeletal muscle, heart, and other fibrous tissues.

Principle

The E.Z.N.A^® Tissue RNA Kits use reversible binding properties of HiBind^® matrix, a new silica-based, time saving spin technology material. HiBind™ matrix avidly, but reversibly, binds RNA under specific conditions allowing proteins and other contaminants to be removed. Nucleic acids are easily eluted with de-ionized water or low salt buffer.

Procedure

The sample is lysed under highly denaturing buffer conditions, such that RNases are inactivated, and the intact Tissue RNA is protected from degradation. After adjusting buffer conditions, samples are treated with Protease K to aid in the degradation and removal of cellular proteins. Cell debris is subsequently pelleted by centrifugation. Ethanol is added to the cleared lysate and the sample is loaded to the E.Z.N.A^® RNA column. With a brief centrifugation or vacuum step, the sample passes through the column and the RNA binds to the Hibind^® matrix. Trace genomic DNA that may co-purified with RNA can be removed by an on column DNase digestion (Catalog # E1091). After two wash steps to remove contaminants, purified total RNA is eluted with RNase free water.

Applications

RNA purified using the E.Z.N.A.^® Tissue RNA Kit can be used in applications such as :

• RT-PCR
• Northern Blot